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目的:制备携抗MUCI单克隆抗体的超声造影剂微泡,并评价其体外寻靶能力。方法:采用异型双功能交联剂将小鼠抗MUCI单克隆抗体与超声造影剂微泡相结合,采用激光粒度仪,扫描电镜及激光共聚焦显微镜评价其粒径,形态及结合率。体外培养小鼠EMT6细胞,将其与靶向微泡相混合,激光共聚焦显微镜评价粘附能力。结果:扫描电镜下显示携抗MUCI单克隆抗体造影剂微泡呈规则球体形态,平均粒径2.88±1.34μm,通过异型双功能交联剂,抗MUC1单克隆抗体可结合至超声造影剂微泡表面,其结合率为77.3±10.4%,靶向造影剂粘附体外培养细胞比例为79.2±13.2%。结论:成功制备携抗MUCI的靶向超声造影剂微泡,并且能很好的识别粘附体外培养乳腺癌细胞。
OBJECTIVE: To prepare ultrasound contrast agent microbubbles against MUCI monoclonal antibody and to evaluate its in vitro targeting ability. METHODS: The mouse anti-MUCI monoclonal antibody was conjugated with ultrasound contrast agent microbubbles using heterobifunctional cross-linking agent. The particle size, morphology and binding rate were evaluated by laser particle sizer, scanning electron microscopy and laser confocal microscopy. Mouse EMT6 cells were cultured in vitro, mixed with the targeted microbubbles, and evaluated for adhesion by laser scanning confocal microscopy. Results: Scanning electron microscopy showed that the anti-MUCI monoclonal antibody-coated microbubbles showed a regular spherical morphology with an average particle diameter of 2.88 ± 1.34μm. The anti-MUC1 monoclonal antibody could bind to the ultrasound contrast agent microbubbles through the heterobifunctional cross-linking agent Surface, the binding rate was 77.3 ± 10.4%, and the ratio of cultured cells adhered by targeted contrast agent was 79.2 ± 13.2%. CONCLUSIONS: Microbubbles targeting anti-MUCI targeted ultrasound contrast agent were successfully prepared and could well recognize the adhesion of breast cancer cells cultured in vitro.